基于HIF/VEGF/Notch信号通路探讨电针“百会”“足三里”对脑缺血大鼠血管新生的影响  

Effect of electroacupuncture at“Baihui”(GV20)and“Zusanli”(ST36)on angiogenesis in the brain of middle cerebral artery occlusion rats based on HIF/VEGF/Notch signaling pathway

作　　者：杨悦悦 吴松 马素娜[1] 关梦雅 王晶莹 任彬彬[2] YANG Yue-yue;WU Song;MA Su-na;GUAN Meng-ya;WANG Jing-ying;REN Bin-bin(Henan University of Chinese Medicine,Zhengzhou 450000,China;The First Affiliated Hospital of Henan University of Chinese Medicine,Zhengzhou 450000)

机构地区：[1]河南中医药大学,郑州450000 [2]河南中医药大学第一附属医院,郑州450000

出　　处：《针刺研究》2024年第10期1030-1039,共10页Acupuncture Research

基　　金：2022年度河南省中医药拔尖人才培养项目(No.2022ZYBJ09);中医药传承与创新人才工程项目(No.CZ0262-10-02);2023年度河南省中医药科学研究专项课题项目(No.2023ZY30010)。

摘　　要：目的:观察电针对脑缺血大鼠缺氧诱导因子(HIF)/血管内皮生长因子(VEGF)/神经源性位点缺口同源蛋白(Notch)信号通路及相关因子的影响,探讨电针对大鼠脑缺血后大脑微血管新生的调控机制。方法:从100只雄性SD大鼠中随机选取15只为假手术组,其余85只大鼠采用改良Zea-Longa法制备左侧大脑中动脉缺血模型,将造模成功的大鼠随机分为模型组、电针组、电针+抑制剂组、抑制剂组,每组15只。电针组大鼠予“百会”、右侧“足三里”电针治疗,疏密波,30 min/次,1次/d;电针+抑制剂组大鼠在电针治疗后,腹腔注射HIF-1α抑制剂YC-1(2.5 mg/kg),1次/d;抑制剂组大鼠腹腔注射HIF-1α抑制剂YC-1(2.5 mg/kg),1次/d。以上各组均连续治疗7 d。采用改良神经功能损伤严重程度量表(mNSS)评分评估大鼠神经功能缺损情况;TTC染色法观察大鼠脑梗死体积百分比;HE染色法观察大鼠缺血半暗带皮层病理学变化;免疫荧光染色法检测CD34表达以计算缺血半暗带皮层微血管密度(MVD);Western blot法和荧光定量PCR法分别检测缺血半暗带皮层脑组织HIF-1α、VEGF、血管内皮生长因子受体2(VEGFR2)、Notch1、Delta样4配体(DLL4)蛋白及mRNA表达水平。结果:与假手术组比较,模型组大鼠mNSS评分、脑梗死体积百分比及脑组织缺血半暗带MVD均升高(P<0.01),缺血侧脑组织HIF-1α、VEGF、VEGFR2、Notch1、DLL4的蛋白及mRNA表达水平升高(P<0.01)。与模型组比较,电针组大鼠mNSS评分、脑梗死体积百分比均降低(P<0.01),脑组织缺血半暗带MVD和缺血侧脑组织HIF-1α、VEGF、VEGFR2、Notch1、DLL4的蛋白及mRNA表达水平均升高(P<0.01);抑制剂组大鼠mNSS评分及脑梗死体积百分比均升高(P<0.01),脑组织缺血半暗带MVD和HIF-1α、VEGF、VEGFR2、Notch1、DLL4的蛋白及mRNA表达水平降低(P<0.05,P<0.01)。与电针+抑制剂组比较,电针组大鼠mNSS评分、脑梗死体积百分比均降低(P<0.01),脑组织缺血半�Objective To observe the effect of electroacupuncture(EA)on hypoxia-inducible factor(HIF)/vascular endothelial growth factor(VEGF)/Notch signaling pathway and related factors in rats with cerebral ischemia(CI),so as to explore its regulatory mechanism underlying improvement of CI by promoting cerebral microangiogenesis.Methods Fifteen male SD rats were randomly selected as the sham surgery(sham)group,while other 85 rats were used to prepare CI model according to the modified Zea-Longa method.The successful CI model rats(n=60)were randomly allocated to model,EA,EA+inhibitor,and inhibitor groups,with 15 rats in each group.EA stimulation(1 Hz/20 Hz,1—2 mA)was applied to“Baihui”(GV20)and right“Zusanli”(ST36)for 30 min,once a day for 7 days.Rats of the 2 inhibitor groups received intraperitoneal injection of YC-1(HIF-1αinhibitor,2.5 mg/kg),once a day for 7 days.The modified neurological severity scale(mNSS,including the motor[muscular state,abnormal movement],sensory[visual,tactile and proprioceptive],balance and reflex functions,0—18 points)was used to assess the rats’neurological deficit state.The percentage of cerebral infarct volume was measured after 2,3,5-triphenyl tetrazolium chloride(TTC)staining,and pathological changes of the ischemic penumbra cortex were observed after H.E.staining.The immunoactivity of CD34 was determined using immunohistochemistry,followed by calculating the microvascular density(MVD)in the ischemic penumbra cortex,and the expression levels of HIF-1α,VEGF,vascular endothelial growth factor receptor 2(VEGFR2),Notch1,and Delta like 4 ligand(DLL4)mRNAs and proteins in the ischemic penumbra of brain tissue were detected using real-time PCR and Western blot,respectively.Results In contrast to the sham group,the mNSS score,percentage of cerebral infarct volume,MVD,and expression levels of HIF-1α,VEGF,VEGFR2,Notch1 and DLL4 proteins and mRNAs were all significantly increased in the model group(P<0.01).In comparison with the model group,the mNSS score and percentage of cerebral i

关 键 词：缺血性脑卒中 电针 缺氧诱导因子/血管内皮生长因子/神经源性位点缺口同源蛋白信号通路 血管新生 

分 类 号：R245[医药卫生—针灸推拿学]